Assessment and reduction of comet assay variation in relation to DNA damage: studies from the European Comet Assay Validation Group

TY - JOUR

T1 - Assessment and reduction of comet assay variation in relation to DNA damage: studies from the European Comet Assay Validation Group

AU - Møller, Peter

AU - Möller, Lennart

AU - Godschalk, Roger W L

AU - Jones, George D D

PY - 2010/3/1

Y1 - 2010/3/1

N2 - The alkaline single cell gel electrophoresis (comet) assay has become a widely used method for the detection of DNA damage and repair in cells and tissues. Still, it has been difficult to compare results from different investigators because of differences in assay conditions and because the data are reported in different units. The European Comet Assay Validation Group (ECVAG) was established for the purpose of validation of the comet assay with respect to measures of DNA damage formation and its repair. The results from this inter-laboratory validation trail showed a large variation in measured level of DNA damage and formamidopyrimidine DNA glycosylase-sensitive sites but the laboratories could detect concentration-dependent relationships in coded samples. Standardization of the results with reference standards decreased the inter-laboratory variation. The ECVAG trail indicates substantial reliability for the measurement of DNA damage by the comet assay but there is still a need for further validation to reduce both assay and inter-laboratory variation.

AB - The alkaline single cell gel electrophoresis (comet) assay has become a widely used method for the detection of DNA damage and repair in cells and tissues. Still, it has been difficult to compare results from different investigators because of differences in assay conditions and because the data are reported in different units. The European Comet Assay Validation Group (ECVAG) was established for the purpose of validation of the comet assay with respect to measures of DNA damage formation and its repair. The results from this inter-laboratory validation trail showed a large variation in measured level of DNA damage and formamidopyrimidine DNA glycosylase-sensitive sites but the laboratories could detect concentration-dependent relationships in coded samples. Standardization of the results with reference standards decreased the inter-laboratory variation. The ECVAG trail indicates substantial reliability for the measurement of DNA damage by the comet assay but there is still a need for further validation to reduce both assay and inter-laboratory variation.

KW - Comet Assay

KW - DNA Damage

KW - DNA-Formamidopyrimidine Glycosylase

KW - Europe

KW - Female

KW - Hela Cells

KW - Humans

KW - Laboratories

KW - Male

KW - Observer Variation

KW - Reference Standards

U2 - 10.1093/mutage/gep067

DO - 10.1093/mutage/gep067

M3 - Journal article

C2 - 20064897

SN - 0267-8357

VL - 25

SP - 109

EP - 111

JO - Mutagenesis

JF - Mutagenesis

IS - 2

ER -